In vivo tumor xenograft model

A mixture containing 100 µL SU-R-786-o cells (5 × 10⁶ cells) or 786-o cells (3 × 10⁶ cells) and 100 µL Matrigel Matrix (Corning, Bedford, MA, USA) was injected subcutaneously into the flanks of female nude mice (BALB/c nu/nu, 6- to 8-weeks-old). (+)-JQ1 (HY-13030; MCE, NJ, USA) was prepared in a vehicle of 10% dimethylsulfoxide and 10% hydroxypropyl-β-cyclodextrin (Sigma, St. Louis, MO, USA) solution and injected at 50 mg/kg intraperitoneally once daily, beginning the day after tumor cell inoculation. The dose was adjusted according to the weight of each mouse, and the volume of injection did not exceed 150 μL. All animal experiments were approved by the animal care review board of Kagoshima University.